Ruscogenin (25D-spirost-5-ene-1(3,3(3-diol; CAS No: 472-11-7), an important plant-derived natural product, exhibits significant anti-tumor, anti-inflammatory, and antioxidant activities. Traditionally, it is produced via inorganic acid hydrolysis, a process that poses serious environmental concerns. Solid acid catalysts, widely applied in hydrolysis, esterification, olefin polymerization, and other chemical transformations, offer a promising alternative to conventional acid-based methods. In this study, a novel strategy was developed by utilizing solid acid catalyst Amberlyst-15 under pressurized conditions for the first time, in which the extract of Ophiopogon japonicus root was hydrolyzed for ruscogenin production. Hydrolysis parameters were optimized using onefactor-at-a-time experiments followed by response surface methodology. Under optimal conditions Amberlyst-15 usage of 0.48 g, hydrolysis temperature of 91 degrees C, and reaction time of 4.5 h - the highest ruscogenin yield of 30.11 % was achieved. Importantly, a high yield maintained at 18.26 % even after seven consecutive reaction cycles of the catalyst for pressurized hydrolysis, well demonstrating its good reusability in this application. The newly established approach was environmental-friendly, offering a green and cost-effective alternative for ruscogenin production.

Oxidative damage is an important cause of aging. Saikosaponin A (SSA) is a major triterpenoid naturally occurring in Radix Bupleuri (RB), a well-recognized superior-grade Chinese herb. Additionally, a rare secondary glycoside, namely prosaikogenin F (PSF) can be obtained after removal of one glucose from SSA. This study aimed to efficiently prepare PSF from SSA, and further investigate and compare their anti-aging activities in Caenorhabditis elegans. Evaluation of antioxidant capacity, heat stress resistance, and aging-related physiological change were conducted, and potential mechanisms were revealed as well. The results showed that Aspergillus niger demonstrated the highest conversion efficiency, achieving a relative PSF yield of 83.9%. Average lifespan of nematodes was extended by 9.32% after treatment with 120 μM SSA, which was further increased to 11.41% while PSF being applied. Under thermal stress stimulation, 120 μM PSF prolonged survival time of nematodes by 35.76% compared to SSA by 33.59%. In addition, they increased the activity of antioxidant enzymes, reduced lipid peroxidation, enhanced the body's resistance to stress damage, and effectively prolonged the lifespan of nematodes. The expression of daf-16, hsp-16.2 and sir-2.1 in the nematodes treated with SSA or PSF were significantly up-regulated, whereas that of the daf-2 gene was significantly down-regulated (p < 0.05). In conclusion, the rare secondary triterpenoid saponin PSF was conveniently obtained from abundant SSA by microbial biotransformation. It showed stronger anti-aging activity than its primary form SSA and acted via regulation of insulin/insulin-like growth factor signaling (IIS) pathway, demonstrating its greater potential for applications in anti-aging and antioxidant.

A butoxy-bridged pyranone-flavonoid hybrid, pyranobutflavin, together with two known compounds (baicalein and baicalin butyl ester), was obtained after high-temperature-little-acid hydrolysis of baicalin naturally occurring in the roots of Scutellaria baicalensis Georgi. Cytotoxicity assays revealed that pyranobutflavin exhibited enhanced anticancer activity compared to baicalin, baicalein, and baicalin butyl ester across various cancer cell lines. Notably, its effect was most pronounced against hepatocellular carcinoma (HCC) cells, with superior selectivity indices over normal cells. Furthermore, this hybrid significantly suppressed proliferation and migration while inducing reactive oxygen species accumulation and apoptosis of HCC cells. Network pharmacology analysis revealed that the mechanism of pyranobutflavin could be closely related to PI3K-Akt and MAPK pathways. Acute toxicity testing established a safe dose (5 mg/kg, i.p.) in mice, at which pyranobutflavin validated its antitumor efficacy in HepG2 xenograft model. These findings identify pyranobutflavin as a promising selective anti-HCC derivative with enhanced therapeutic potential over its natural precursors.

Thrombosis triggers various severe diseases, while antithrombotic drugs carry bleeding risks, making the development of novel natural anticoagulants a subject of widespread attention. Poecilobdella manillensis, a prevalent medicinal leech, exhibits remarkable anticoagulant and antithrombotic activities. However, the material basis underlying its anticoagulant effects remains insufficiently investigated. This study aims to mine anticoagulant peptides from P. manillensis by peptidomics analysis, elucidate the material basis of its anticoagulant activity, and provide candidate molecules for developing novel natural anticoagulant drugs. Proteins extracted from P. manillensis were enzymatically digested and fractionated using DEAE-52 and CN columns. The resulting peptide components were analyzed by UPLC-Q-Orbitrap HRMS, and peptide sequences were matched against proteomic databases using Proteome Discoverer. Anticoagulant peptides were predicted using the BIOPEP-UWM database and PeptideRanker server, followed by in vitro and in vivo activity validation. Results showed that the hydrolysate consisted predominantly of low-molecular-weight peptides. 1533 peptides with Mw < 3000 Da (length < 20 amino acids) were identified from the PM-A2 and PM-A3 fractions, accounting for 40.76

ABSTRACTIntense pulsed light (IPL) is an efficient sterilization technology for instantaneous intense UV predominantly, while the decontamination was weakened on green Sichuan pepper (GSP) for “shadow effect.” Catalytic infrared (CIR) has excellent decontamination on uneven surfaces for long‐wave diffraction while was prone to cause flavor decline for “thermal effect.” In this study, a combined method of CIR and IPL was attempted to settle the dispute of decontamination effect and flavor decline on GSP. The inactivation kinetics on the change of bacterial enumeration over sterilization time were determined, and the results showed the sterilization effect of IPL on GSP could be improved significantly after CIR pretreatment, which was promoted from 0.65 ± 0.07 to 2.62 ± 0.07 Lg (p < 0.05), and could realize the active bacteria on commercial GSP was below 100 CFU g−1. Moreover, the flavor of GSP was measured on color and aroma changes, volatile oil content, and total alkaloid content, and the antioxidant function was measured on total reducing power, ABTS+•, and •OH− clearance rates. The results showed CIR could give rise to color change and volatile oil decrease of GSP, which indicated that the treatment time of CIR should be restricted in 5 min based on relevant Chinese national standard. CIR and IPL showed a difference to several oxygen free radicals, and the impact could be weakened by a combined method. In conclusion, under the same sterilization rate, CIR pretreatment could promote the decontamination of IPL on GSP and could weaken the influence of CIR or IPL on the quality of GSP.

Intense pulsed light (IPL) is an emerging surface antimicrobial technology characterized by prominent efficiency but the performance in the decontamination of granular foods is yet to be improved. Using S. Enteritidis as a model bacterium, this article attempted to resolve the confusion on bactericidal mechanism of IPL treatment on spice products. The bactericidal effects of IPL treatment were compared on spices with different surfaces, and the result indicated that "shadow effect" from the shorter UV wavelengths of IPL was one of the reasons for decrease in antimicrobial efficiency. Then the critical bactericidal mechanism of IPL has been investigated by the evaluation of the viable but non-culturable (VBNC) state of the bacteria, apoptosis under IPL stress, cytomembrane destruction and DNA damage. The results revealed that "photochemical effect" of UV light and "photothermal effect" of IR light could have played different roles. After IPL treatment of low dosage (fluence <3.12 J/cm2), the UV light damaged the intracellular DNA and turned the bacteria into their VBNC state, which has caused an overestimate of bactericidal effect. On the other hand, the IR light inhibited bacterial resuscitation by destruction of the cytomembrane when IPL treatment of high dosage (fluence >6.24 J/cm2) was applied on green Sichuan pepper (GSP) with uneven surface. As a consequence, "photothermal effect" and diffraction of IR light improved the decontamination effect of IPL treatment against S. enteritidis, and higher dosage irritation could be a preferred selection for the shelf-life extension of spice products.

Human milk oligosaccharides (HMOs) are a diverse group of lactose-derived carbohydrates and represent the third most abundant component in human milk. Fucosylated HMOs, which are particularly abundant, offer significant benefits to infants and have garnered increasing interest in recent decades. However, their widespread application in the food industry and other sectors remains limited due to challenges associated with their synthesis and isolation on a large scale. This review summarizes the chemical structures of fucosylated HMOs and their biological functions, critically evaluates conventional and cutting-edge synthetic strategies, and discusses the major challenges and future prospects in this field. Despite advancements, our understanding of the bioactivities and functions of fucosylated HMOs remains limited, necessitating further in vitro and in vivo studies. Fucosidase-catalyzed transfucosylation and metabolic engineering have emerged as promising strategies for their synthesis. Additionally, plant-based synthesis offers a sustainable alternative for large-scale production. Genetic modification, reaction condition optimization, and protein engineering have significantly improved the production of fucosylated HMOs. However, challenges such as identifying economical substrates, obtaining enzyme crystal structures for molecular engineering, developing advanced purification techniques, and ensuring the safety of synthesized products remain. Systematic evaluation and integration of knowledge are essential to drive progress in this field.

Diosgenin (25R-spirost-en-3 beta-ol; CAS No.: 512-04-9), a plant-derived natural product, has significant importance for manufacturing steroid-based drugs. It is primarily prepared by direct acid hydrolysis, but this out-of-date process is not environmentally friendly. In recent decades, ionic liquids have shown good potential to replace conventional organic solvents in many fields. The aim of this study was to develop a novel approach for diosgenin production, in which the acidic ionic liquid [BHSO(3)MIm]HSO4 was employed under pressurised conditions to hydrolyse the crude saponin of Dioscorea zingiberensis C. H. Wright tubers for the first time. The hydrolysis conditions were optimised through a one-factor-at-a-time experiment, and the maximum yield of diosgenin was achieved at 3.71 +/- 0.18 % with an ionic liquid concentration of 0.5 M, a solid-liquid ratio of 1:30 g/mL, a hydrolysis temperature of 140 degrees C, and a hydrolysis duration of 0.5 h. The diosgenin yield was 75.67 % of the maximum yield after six consecutive applications of [BHSO(3)MIm]HSO4. The yield achieved was comparable to pressurised acid hydrolysis and microwave-assisted ionic liquid hydrolysis and significantly higher than that of direct acid hydrolysis (P<0.01). Meanwhile, catalyst consumption was 0.123 mol/g diosgenin, which was much lower than that of microwave-assisted ionic liquid hydrolysis (0.600 mol/g). Moreover, the hydrolysis reaction was completed within 0.5 h, which was only one-third of the conventional pressurized acid hydrolysis. This newly established method has significant merit in hydrolysis duration and is associated with decreased catalyst consumption, and [BHSO(3)MIm]HSO4 is of good reusability, making it a greener and more economical method for diosgenin preparation.

Baicalein, usually obtained after the removal of 7-O-beta-D-glucuronic acid from baicalin, has strong pharmacological activity. In this study, a novel strategy for efficient and eco-friendly preparation of baicalein was established by high-temperature-little-acid hydrolysis of baicalin. The effects of inorganic acids, solvents, solid-liquid ratio, HCl concentration and hydrolysis duration on the yield of baicalein were investigated by one-factor-at-a-time experiment. Then, an orthogonal experiment design was conducted for the optimal hydrolysis conditions, namely, baicalin was boiled in n-BuOH solution containing 0.50% HCl (w/v) at 130 degrees C for 2.0 h with solid-liquid ratio at 1:25 (g/mL). In the 60-fold scale-up experiment, the yield of baicalein was achieved at 83.5% +/- 6.3%, and its purity was increased from 74.2% to 98.7% after purification. Compared with reported methods, acid consumption per unit product of the newly developed strategy was approximately 80 times lower than that of conventional acid hydrolysis, and time consumption was substantially reduced. Overall, the novel strategy showed great prospects for a cleaner large-scale production of baicalein, which provides a new alternative desirable for the preparation of natural products with high efficiency and low consumption in the pharmaceutical industry.

Leeches are a well-known animal-derived health supplement commonly used as an anticoagulation and antithrombosis agent; however, adulteration and counterfeiting are often made for illegal profits. To identify leech species, this study developed a rapid, simple, and visualized method based on loop-mediated isothermal amplification (LAMP), which relies on a specific primer set designed according to the mitochondrial DNA control region of the target species. Quantitative polymerase chain reaction (qPCR) was also employed in parallel to compare the sensitivity and confirm the primer specificity. Primer sets with high specificity were successfully screened for LAMP reactions against four common leech species on the market. All of them have produced typical amplification profiles of the target sequences in qPCR reactions with significantly lower amplification sensitivity than LAMP assay. The newly established LAMP method in this study can be accomplished within 1 h, and it could be successfully applied for on-site visual identification of mislabeling and adulteration in the leech market.

Isoquercitrin has superior in vivo bioactivities with respect to its primary glycoside rutin. Its conventional preparation was ineffective, with large chemical consumption and many by-products. Rhamnose, a high value-added monosaccharide, is usually separated from acid hydrolytes of rutin. This study aimed to establish a novel enzymatic hydrolysis-based approach for their preparation. α-L-rhamnosidase was expressed in Pichia pastoris GS115 and applied to enzymolysis of rutin. Then, one-factor-at-a-time optimisation of hydrolysis conditions was performed. Two compounds were produced in 0.02 M HAc-NaAc buffer (pH4.50) containing α-L-rhamnosidase/rutin (1:4, w/w) at 60 °C. Consequently, 20.0 g/L rutin was completely hydrolysed in 2 hrs, and isoquercitrin was obtained after purification by HPD-100 resin. Additionally, rhamnose was enriched by decolorisation and crystallisation. MD simulation analysis suggested that rutin was catalysed on the hydrophobic surface of r-Rha1 with van-der-Waals force being main driving force. This strategy is an efficient approach for preparation of isoquercitrin and rhamnose.

目的 建立特异性引物扩增鉴定金嗓清音丸中僵蚕成分.方法 根据家蚕、白僵菌、绿僵菌线粒体全基因序列差异设计特异性引物PM2、PB2、PA2,采用十六烷基三甲基溴化铵法、酚仿抽提法提取纯化总DNA,并以此为模板进行特异性引物扩增和方法学考察,产物以2％琼脂糖凝胶电泳检测.结果 家蚕、白僵菌、绿僵菌DNA经PCR扩增后,分别在82、94、144 bp处产生条带,并且未观察到非特异性扩增条带.PM2、PB2、PA2对相应DNA的检测灵敏度分别为0.10、0.10、0.010 ng/μL.来自同一厂家的6批样品中同时检出家蚕、白僵菌DNA,但未检出绿僵菌DNA,而另外2批样品仅检出家蚕DNA.结论 该方法方便可靠,专属性强,有助于更好地监控金嗓清音丸等含僵蚕中药成方制剂的质量,可为其临床用药的安全性和有效性提供保障,并为其他中成药中动物药的专属性鉴定提供参考.

A combined method of catalytic infrared (CIR) and intense pulsed light (PL) was investigated to settle a dispute of bacteriostasis on green Sichuan pepper (GSP), and the synergistic principle was elucidated from several aspects. The synergistic bacteriostasis was confirmed first on GSP by the plate counting method. On bacterial cells, leakage of cytoplasmic contents, intracellular protein and DNA breakages were investigated, and the difference between CIR and PL indicated the possibility of synergism. On a single bacterium, the expected synergy was not achieved on the plate but was obtained on GSP, and the bacterial cells treated by the combined method were mainly broken under SEM, indicating that the alliance technology could increase the damage of PL to bacterial cells on the rough surface of GSP. On multiple bacteria, synergy was obtained similarly, and the bacterial community on GSP was investigated with high-throughput sequencing. The community composition on GSP could be changed by the treatments. In conclusion, synergistic bacteriostasis on GSP was achieved by a combined method of CIR and PL, originating from a complementary effect on the microbiome and a promotion of bacteriostatic capacity on rough surfaces. These findings are expected to solve the bacteriostatic difficulty of spices with rough surfaces and have important implications for the development of effective bacteriostatic strategies in food preservation.

Prosapogenin A is a secondary saponin in Dioscorea zingiberensis, and it showed remarkable pharmacological effects. Due to very low content and lack of well-developed biotransformation, its preparation was not efficient and clean. This study aims to establish an eco-friendly strategy for preparation of Prosapogenin A from plant material. Physical separation was employed to recycle starch and cellulose, and then D101 resin and polyamide packed-bed column was incorporated for purification of total steroidal saponins (TSS). After these pretreatments, purity of TSS was largely increased to 83.2% with recovery at 87.6%, which was subjected to enzymatic hydrolysis. Optimized reaction system was constructed in 0.20 M HAc-NaAc buffer (pH4.2) containing cellulase/TSS (3:1, w/w), and the hydrolysis was performed at 53 degrees C for 6 h. Consequently, TSS was almost completely hydrolyzed to Prosapogenin A, while the highest yield reached 5.62%. The newly proposed approach is promising for efficient preparation of Prosapogenin A in industrial applications.

Background Prosaikogenin F (PSF) has stronger anti-cancer bioactivity than saikosaponin A (SSA), however, it was hardly isolated due to its trace amount in the raw material of Radix Bupleuri (RB). In addition, the active chemical constituent was unstable under acidic conditions owing to a 13,28-epoxy-ether moiety at the D ring of its aglycone. Objectives This study was to develop an appropriate method for obtaining acid-sensitive PSF from SSA abundant in RB. Materials and Methods Enzymatic hydrolysis was employed and snailase was selected due to its good hydrolysis performance under nearly neutral circumstances. Hydrolysis conditions were then optimized by one-factor-at-a-time experimentation before response surface methodology (RSM) by Box-Behnken Design (BBD). Results The reaction system was constructed in Na 2 HPO 4 -NaH 2 PO 4 buffer (pH 6.0) containing snailase/SSA (44:1) at 39°C, then the hydrolysis lasted for 12 h. Therefore, the highest conversion ratio of SSA was achieved at 100.0%. Conclusion The newly proposed method is eco-friendly for obtaining acid-sensitive PSF, which lays a solid foundation for its development to be an anti-cancer new drug.

Abundant starch was isolated from Dioscorea zingiberensis C.H. Wright, a novel and underutilized industrial crop resource. In this study, an intelligent packaging film able to indicate food freshness was developed and characterized. D. zingiberensis starch (DZS) was bleached first, and its particle size, total starch content, amylose content, and gelatinization temperature were then measured. Butterfly pea (Clitoria ternatea Linn.) flowers were selected as the source of polyphenols, which rendered the prepared film intelligent and progressively blue-violet. SEM and FT-IR analyses showed the homogeneous dispersion of butterfly pea flower extract (BPE) in the film. The BPE-loaded film showed improved flexibility and resistance to UV and oxidation while maintaining sufficient mechanical strength and physical properties. Moreover, the film underwent a distinguishable color change from red to blue-violet and finally to green-yellow with increasing pH from 2 to 13. Similar color alteration also occurred when the film was exposed to ammonia. When the film was used to monitor the freshness of chicken stored at room temperature, it exhibited an obvious color change, implying its deterioration. Therefore, the newly developed BPE-DZS film, which was produced from readily accessible natural substances, can serve as an intelligent packaging material, indicating food freshness and prolonging shelf life.

Improving physiological activity of primary ginsenosides through biotransformation is of great significance for food applications. In this study, gynostapenoside XVII, gynostapenoside LXXV, ginsenoside F2, and ginsenoside CK were obtained by enzymolysis of an accessible extract composed of ginsenoside Rb1 and Rd. Their effects on melanin content and tyrosinase activity were compared in vitro, and molecular docking simulation was employed to elucidate the interaction between tyrosinase and individual saponin. The results indicated that four rare ginsenosides decreased tyrosinase activity, melanin content and microphthalmia-associated transcription factor (MITF) expression level, more greatly than their primary ginsenosides, and they were more readily to bind with ASP10 and GLY68 at active site of tyrosinase to inhibit tyrosinase activity as well. These findings suggested that the rare ginsenosides obtained by enzymolysis had excellent anti-melanogenic effect, which could expand the application of ginsenosides in the field of functional foods and health supplements.

In this study, abundant starch was separated from the industrial crop Dioscorea zingiberensis C.H. Wright (DZW), and a novel bioactive packaging film loaded with oregano essential oil (OEO) was prepared and characterized. NaClO solution worked as a bleacher to prepare uniform starch powder from DZW tubers. OEO was selected from among three essential oils of Labiatae family plants for its strongest antibacterial activity. After the addition of OEO into the starch-based film, the UV-vis shielding property and antioxidant activity were enhanced. Meanwhile, the films still have a considerable performance in transparency, mechanical strength and water vapor permeability after incorporated with OEO. Furthermore, the 3% OEO-loaded starch film exhibited the strongest antibacterial activity against Bacillus subtilis, Escherichia coli and Staphylococcus aureus. It effectively lowered the total viable count of fresh chicken under 4 degrees C preservation conditions. These results revealed that the OEOloaded DZW starch film can exert a positive effect on maintaining the quality and extending the shelf life of fresh meat. Therefore, readily accessible DZW tubers and oregano are very promising resources for application in degradable bioactive packaging film.

Catalytic infrared (CIR) technology for sterilization has not been widely applied owing to the lack of comprehensive and in-depth mechanistic research. This study aims to explore further the inactivation mechanism of CIR against Pseudomonas aeruginosa in green Sichuan pepper (GSP). First, the working pattern of the CIR was investigated using a colony counting method in different layouts. The results indicated that the bacteriostatic effect on GSP was weaker than that on plates with low-dose CIR radiation, and both the surrounding air and surface moisture content affected the sterilization rate of P. aeruginosa. Second, the bacteriostatic mechanism of CIR against P. aeruginosa was investigated. The results indicated that the bacteriostatic effect of CIR could be mainly caused by the destruction of the cell membrane, inhibition of swimming motility, and membrane dysfunction simultaneously, and it was also caused by weaker DNA breakage compared with pulse strong light. Finally, the flavour influence of CIR in GSP was determined in terms of volatile oil content, aroma component, alkaloid content, and colour change. After 0-4 min of CIR treatment, the changes in the flavour of GSP were all within the acceptable range. Consequently, CIR technology is a promising approach for effective decontamination of the GSP surface, which works mainly on cell cover and can weakly penetrate the cell membrane.

Prosapogenin A (also known as Progenin III and polyphyllin V), a secondary steroidal saponin in Dioscorea zingiberensis tubers, has more superior pharmacological activities than its primary form Protogracillin. Conventional preparation methods, such as column chromatography, and acidic hydrolysis, are of very low efficiency and limited scale due to tedious procedures and large consumption of organic solvent. This study aims to establish a convenient method for efficiently obtaining Prosapogenin A by enzymatic hydrolysis of abundant Protogracillin in the raw material. In light of the highest hydrolysis performance, β-dextranase was selected from four commercial enzymes in this application. After optimization of the conditions by the response surface methodology using Box–Behnken design, the enzymatic hydrolysis was carried out in 0.20 M HAc-NaAc buffer (pH 4.81) containing β-dextranase/Protogracillin (5.0:1, w/w), and the system was constantly kept at 56.7°C water bath for 4 h. Consequently, Protogracillin has been almost completely hydrolyzed to be Prosapogenin A and the highest yield was 96.4 ± 1.4%. The newly proposed approach is efficient and promising for conveniently obtaining Prosapogenin A and aimed to provide a laboratory-scale experimental foundation for the preparation of Prosapogenin A in industrial applications.